Journal: International Journal of Molecular Medicine

Article Title: Ebastine targets HER2/HER3 signaling and cancer stem cell traits to overcome trastuzumab resistance in HER2-positive breast cancer

doi: 10.3892/ijmm.2026.5751

Figure Lengend Snippet: EBA impairs cancer stem cell-like properties. (A) BT474 and SKBR3 cells were treated with EBA for 48 h, and ALDH1 activity was assessed by flow cytometry using the Aldefluor assay. DEAB was used to define the baseline of Aldefluor-positive fluorescence. (B) BT474 cells (5x10 4 cells/ml) were plated in ultra-low attachment dishes and cultured in the presence or absence of EBA for 5 days. The number and volume of mammospheres were measured by microscopy. (C) Overall survival of patients with breast cancer stratified by the co-expression of ALDH1A1 and CD44. (D) Spearman correlation analysis of ALDH1A1 and CD44 mRNA levels in patients with HER2-positive breast cancer from The Cancer Genome Atlas cohort (n=76). Kaplan-Meier survival analyses of patients with HER2-overexpressing breast cancer stratified by (E) ALDH1A1 and (F) CD44 expression. Patients were divided into high- and low-expression groups based on the median gene expression. Statistical significance was determined using the log-rank test. (G) JIMT-1 cells were treated with EBA (3 μ M) for 48 h and the CD44 high /CD24 low cell populations were identified by flow cytometry. (H) JIMT-1 cells (1.5x10 4 cells/ml) were cultured under serum-free suspension conditions in the presence of EBA (3 μ M) for 8 days. Mammosphere number and volumes were quantified. ** P<0.01 and **** P<0.0001 vs. vehicle-treated control (0 μ M EBA). EBA, ebastine; ALDH, aldehyde dehydrogenase; DEAB, diethylaminobenzaldehyde; CTL, control; ISO, isotype.

Article Snippet: The human breast cancer cell lines SKBR3, BT474, MDA-MB-453 (American Type Culture Collection) and JIMT-1 (Leibnitz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH) were cultured in DMEM, MEM or RPMI-1640 (all Sigma-Aldrich; Merck KGaA) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and 100 U/ml penicillin-streptomycin at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Activity Assay, Flow Cytometry, Fluorescence, Cell Culture, Microscopy, Expressing, Gene Expression, Suspension, Control

Journal: Breast Cancer Research : BCR

Article Title: High levels of circulating miR-19a-3p in patients with metastatic HER2 + breast cancer are associated with a favorable prognosis and anti-tumor immune responses

doi: 10.1186/s13058-025-02174-8

Figure Lengend Snippet: Expression and secretion levels of miR-19a-3p in breast cancer cells. a) Breast cancer cells express different levels of miR-19a-3p depending on the subtypes, with TNBC (basal) expressing the highest levels compared to luminal B, PAM50 Normal, HER2+, luminal A, and normal cells (TCGA data). The data are shown as box and whisker plots, where the box encompasses the 25th percentile, median, and 75th percentile, and the whiskers extend to 1.5 times the interquartile range; b) the two HER2+ breast cancer cell lines KPL-4 and SKBR3 express higher level of miR-19a-3p than the luminal A MCF-7, consistent with TCGA data; c) the three breast cancer cell lines secrete varying amounts of miR-19a-3p, following an expression pattern similar to that in their parental cells. The results are shown as the mean ± SD of three technical replicates from 3 experimental replicates

Article Snippet: MCF-7 (ER +) and SKBR3 (HER2 +) cells were obtained from the American Type Culture Collection (Manassas, VA, USA), and KPL-4 (HER2 +) cells were provided courtesy of Dr. Junichi Kurebayashi (Kawasaki Medical School, Kurashiki, Japan) [ ].

Techniques: Expressing, Whisker Assay

Journal: Breast Cancer Research : BCR

Article Title: High levels of circulating miR-19a-3p in patients with metastatic HER2 + breast cancer are associated with a favorable prognosis and anti-tumor immune responses

doi: 10.1186/s13058-025-02174-8

Figure Lengend Snippet: Central memory T (T CM ) cell phenotype of CD4 + Th1 cells and immune cell populations in the peripheral blood of patients with metastatic HER2 + breast cancer. a-b) CD4 + Th1 cells expressing high levels of miR-19a-3p developed a central memory T (T CM ) cell phenotype (CD45RO + CCR7 + CD62L +) by the end of the cell culture; c-e) in a small cohort of patients with metastatic HER2 + breast cancer, those with a good prognosis and high serum levels of miR-19a-3p showed a trend of higher percentage of CD4 + Th1 than CD4 + Th2 cells (ratio: IFN-γ + /IL-4 +), although not statistically significant (p = 0.08130) and activated CD4 + T and NK cells circulating in their blood. The results of panel a are shown as the mean ± SD of 3 technical replicates. The results in panel b are presented as the most representative of 3 technical replicates and two experimental replicates. The results in panels c-e are shown as the median with a 95% confidence interval (CI)(Available data from n = 15)

Article Snippet: MCF-7 (ER +) and SKBR3 (HER2 +) cells were obtained from the American Type Culture Collection (Manassas, VA, USA), and KPL-4 (HER2 +) cells were provided courtesy of Dr. Junichi Kurebayashi (Kawasaki Medical School, Kurashiki, Japan) [ ].

Techniques: Expressing, Cell Culture

Journal: Breast Cancer Research : BCR

Article Title: High levels of circulating miR-19a-3p in patients with metastatic HER2 + breast cancer are associated with a favorable prognosis and anti-tumor immune responses

doi: 10.1186/s13058-025-02174-8

Figure Lengend Snippet: Proposed model of TME of HER2 + breast cancer modulated by NK cell-mediated ADCC. The proposed model is based on findings from our current and previous work. Further studies are needed to validate and confirm our hypothetical model. Trastuzumab binds to HER2 + cells and engages CD16 on NK cells, activating them and inducing ADCC. This leads to perforin/granzyme B-mediated apoptosis and death of HER2 + breast cancer cells, as well as the release of miR-19a-3p into the TME and eventually into the bloodstream. ADCCalso stimulates the production and secretion of IFN-γ and T cell-recruiting chemokines from NK cells, which activate DCs, promote their migration to draining lymph nodes, and induce migration of lymphocytes into the TME. This process enables TAA cross-presentation to T cells (both CD4 + and CD8 +) and triggers the production and secretion of IL-12 by DCs. IL-12 and IFN-γ promote the differentiation of CD4 + and CD8 + T cells into CD4 + Th1 cells and CD8 + CTLs and enhance the cytotoxic activity of CD8 + CTLs and NK cells. IL-2 promotes the proliferation of CD4 + Th1 cells, CD8 + CTLs, and NK cells, while IFN-γ from CD4 + Th1 and NK cells further activates DCs (enhancing TAA cross-presentation and cytokine release). The activation and differentiation of CD4 + Th1 cells increase the expression and secretion of miR-19a-3p into the TME and bloodstream. Activated CD8 + CTLs infiltrate tumor tissue and work with NK cells to kill tumor cells, further increasing the release of miR-19a-3p. Additionally, CD4 + Th1 cells differentiate into CD4 + Th1 T CM cells (CD45RO + CCR7 + CD62L +). TAAs from the TME and presented by DCs can maintain the expression and secretion of miR-19a-3p by stimulating CD4 + Th1 T CM cells in draining lymph nodes and contributing to higher serum levels of miR-19a-3p. (This figure was created in BioRender )

Article Snippet: MCF-7 (ER +) and SKBR3 (HER2 +) cells were obtained from the American Type Culture Collection (Manassas, VA, USA), and KPL-4 (HER2 +) cells were provided courtesy of Dr. Junichi Kurebayashi (Kawasaki Medical School, Kurashiki, Japan) [ ].

Techniques: Migration, Activity Assay, Activation Assay, Expressing

Journal: bioRxiv

Article Title: Ionic Regulation of Mechanosurveillance and Metastasis via the MRTFA/KCNMB1 Axis

doi: 10.64898/2026.01.13.699089

Figure Lengend Snippet: (A) Venn diagram and gene ontology analyses of the SRF independent genes that are exclusively upregulated by MRTFA Y238A in both AT-3 and E0771 cells to at least 2-fold or more. See Table S2 for full results. (B) Examples of statistically significant GSEA for MRTFA high/low breast cancer patient tumors from TCGA. (C) Examples of statistically significant GSEA for SRF high/low breast cancer patient tumors from TCGA. (D) Relative mRNA expression levels of MRTFA and MRTFB in shMRTFA/B and SRF in shSRF mouse cell lines. Data are shown as fold change calculated from 3 technical replicates per sample and normalized to GAPDH levels. Each graph is representative of 3 independent experiments. (E) Relative mRNA expression levels of MRTFA and MRTFB in shMRTFA/B and SRF in shSRF human cell lines. Data are shown as fold change calculated from 3 technical replicates per sample and normalized to GAPDH levels. Each graph is representative of 3 independent experiments. (F) Relative mRNA expression levels of canonical YAP target gene CCN2, which encodes connective tissue growth factor (CTGF), and KCNMB1 in AT-3 cells overexpressing wildtype YAP or hyperactive YAP 5SA mutant or treated for 24 hours with TGFβ inhibitor SB-505124. (G) KCNMB1 human and mouse genetic loci analyzed for SRF (purple tracks) binding by using University of California Santa Cruz Genome Browser data publicly available Chromatin immunoprecipitation sequencing data. Each purple solid line under the ReMap density map indicates an independent dataset where SRF was found to be associated with the locus. In the Encode cCREs track, red solid boxes indicate promoter regions, orange and yellow solid boxes indicate proximal and distal enhancers, respectively, and blue tracks indicate CTCF only binding regions. (H) Chromatin immunoprecipitation-PCR from AT-3 cells using KCNMB1 primers and the indicated antibodies against SRF, MRTFA, histone H3 (as a positive control) and IgG (negative control). Water was used as input material as a negative control for the PCR reaction. Image is representative of 3 independent experiments. (I) Raw counts from bulk RNA-seq data for Myocd , Mrtfa , and Mrtfb genes in AT-3 and E0771 cells.

Article Snippet: SKBR3 cells were cultured in McCoy’s 5A media (ATCC #30-2007) supplemented with 10% (vol/vol) FBS.

Techniques: Expressing, Mutagenesis, Binding Assay, ChIP-sequencing, Chromatin Immunoprecipitation, Positive Control, Negative Control, RNA Sequencing